The ceramide-1-phosphate analogue PCERA-1 modulates tumour necrosis factor-α and interleukin-10 production in macrophages via the cAMP-PKA-CREB pathway in a GTP-dependent manner

Dorit Avni, Amir Philosoph, Michael M. Meijler, Tsaffrir Zor*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Summary The synthetic phospho-ceramide analogue-1 (PCERA-1) down-regulates production of the pro-inflammatory cytokine tumour necrosis factor-α (TNF-α) and up-regulates production of the anti-inflammatory cytokine interleukin-10 (IL-10) in lipopolysaccharide (LPS) -stimulated macrophages. We have previously reported that PCERA-1 increases cyclic adenosine monophosphate (cAMP) levels. The objective of this study was to delineate the signalling pathway leading from PCERA-1 via cAMP to modulation of TNF-α and IL-10 production. We show here that PCERA-1 elevates intra-cellular cAMP level in a guanosine triphosphate-dependent manner in RAW264.7 macrophages. The cell-permeable dibutyryl cAMP was able to mimic the effects of PCERA-1 on cytokine production, whereas 8-chloro-phenylthio-methyladenosine-cAMP, which specifically activates the exchange protein directly activated by cAMP (EPAC) but not protein kinase A (PKA), failed to mimic PCERA-1 activities. Consistently, the PKA inhibitor H89 efficiently blocked PCERA-1-driven cytokine modulation as well as PCERA-1-stimulated phosphorylation of cAMP response element binding protein (CREB) on Ser-133. Finally, PCERA-1 activated cAMP-responsive transcription of a luciferase reporter, in synergism with the phosphodiesterase (PDE)-4 inhibitor rolipram. Our results suggest that PCERA-1 activates a Gs protein-coupled receptor, leading to elevation of cAMP, which acts via the PKA-CREB pathway to promote TNF-α suppression and IL-10 induction in LPS-stimulated macrophages. Identification of the PCERA-1 receptor is expected to set up a new target for development of novel anti-inflammatory drugs.

Original languageEnglish
Pages (from-to)375-385
Number of pages11
JournalImmunology
Volume129
Issue number3
DOIs
StatePublished - Mar 2010

Keywords

  • Cyclic adenosine monophosphate response element binding protein
  • Inflammation
  • Interleukin-10
  • Lipopolysaccharide
  • Macrophages
  • Tumour necrosis factor-α

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